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Kinetics of Dodecanedioic Acid and Effect of Its Administration on Glucose Kinetics in Rats by ALESSANDRO BERTUZZI', GELTRUDE MINGRONE', ANDREA DE GAETAN03

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Kinetics of Dodecanedioic Acid and Effect of Its Administration on Glucose Kinetics in Rats by ALESSANDRO BERTUZZI', GELTRUDE MINGRONE', ANDREA DE GAETAN03 Downloaded from British Journal of Nutrition (1997), 78, 143-153 143 https://www.cambridge.org/core Kinetics of dodecanedioic acid and effect of its administration on glucose kinetics in rats BY ALESSANDRO BERTUZZI', GELTRUDE MINGRONE', ANDREA DE GAETAN03, ALBERT0 GANDOLH', ALDO V. GREC02 AND SERENELLA SALINAR14 . IP address: 'Istituto di Analisi dei Sistemi ed Informatica del CNR, Viale Manzoni 30, 00185 Roma, Italy 21stituto di Clinica Medica, Universita Cattolica del Sacro Cuore, Largo A. Gemelli 8, 00168 Roma, Italy 3Centro di Studio per la Fisiopatologia dell0 Shock del CNR, Universita Cattolica del Sacro Cuore, 170.106.33.42 Largo A. Gemelli 8, 00168 Roma, Italy 4Dipartimento di Informatica e Sistemistica, Universita di Roma 'La Sapienza', Roma, Italy , on (Received 30 April 1996 Revised 26 September 1996 -Accepted 17 October 1996) - 28 Sep 2021 at 04:31:38 Dodecanedioic acid (C12), a saturated aliphatic dicarboxylic acid with twelve C atoms, was given as an intraperitoneal bolus to male Wistar rats, with the aim of evaluating C12 suitability as an energy substrate for parenteral nutrition. The 24 h urinary excretion of C12 was 3.9 % of the administered dose. C12 kinetics were investigated by a one-compartment model with saturable tissue uptake and reversible binding to plasma albumin. The analysis of plasma concentration and urinary excretion , subject to the Cambridge Core terms of use, available at data from different animals yielded the population means of the kinetic parameters: renal clearance was 0.72ml/min per kg body weight (BW) (much smaller than inulin clearance in the rat), and maximal tissue uptake was 17.8 pmol/min per kg BW corresponding to 123.7J/min per kg BW. These results encourage the consideration of C12 as a possible substrate for parenteral nutrition. To investigate the effect of C12 administration on glucose kinetics, two other groups of rats, one treated with an intraperitoneal bolus of C12 and the other with saline, were subsequently given an intravenous injection of D-[U-'4C]glucose in a tracer amount. Radioactivity data of both control and C12-treated rats were analysed by means of a two-compartment kinetic model which takes into account glucose recycling. The estimates of glucose pool size (2.3 mmol/kg BW) and total-body rate of disappearance (82.1 pmol/min per kg BW) in control rats agreed with published values. In C12- treated rats, the rate of disappearance appeared to be reduced to 36.7 pmol/min per kg BW and the extent of recycling appeared to be negligible. Dodecanedioic acid: Glucose: Parenteral nutrition https://www.cambridge.org/core/terms Many studies have been performed on the synthesis and fate of dicarboxylic acids (DA) in living organisms since the paper of Verkade & Van der Lee (1934). These authors showed that, after the administration of triundecylin to two healthy volunteers, a considerable amount of undecanedioic acid, the corresponding dicarboxylic acid with eleven C atoms, was excreted in the urine. Therefore, they hypothesized that the terminal methyl group of triundecylin underwent oxidation leading to the formation of a carboxylic group. Subse- quently, the authors called this kind of oxidation o-oxidation. Although the results on the quantitative importance of o-oxidation in the oxidation of fatty acids appear to be quite . confusing (Wada & Usami, 1977; Bjorkhem, 1978), there is consensus on the fact that https://doi.org/10.1079/BJN19970126 when the b-oxidation route is impaired, o-oxidation becomes a very important metabolic pathway for oxidation of fatty acids (Lindstedt et al. 1976; Mortensen & Gregersen, 1981). Dicarboxylic acids with chain lengths shorter than or equal to twelve C atoms are b- oxidized in both mitochondria and peroxisomes (Pettersen & Aas, 1973; Leighton et al. Downloaded from 144 A. BERTUZZI ET AL. https://www.cambridge.org/core 1989). Odd-chain DA are split to acetyl-CoA and, as a terminal product, to malonic acid, which is not further oxidizable and is the starter of fatty acid synthesis. In contrast, even- chain DA can be completely oxidized, since they produce acetyl-CoA and succinyl-CoA which are completely oxidized in the Krebs cycle. In addition, succinyl-CoA is a gluconeogenic substrate (Wada & Usami, 1977; Kou & Tserng Shiow-Jen, 1991). DA have been investigated as possible alternative fuel substrates for parenteral nutrition (Mingrone & et al. 1989, 1991; Bertuzzi et al. 1991; Greco Mingrone, 1995). IP address: Previously, we reported that the continuous intravenous infusion of disodium sebacate (a DA with ten C atoms) caused a significant reduction of glucose uptake in normal and obese individuals and subjects with insulin-dependent diabetes mellitus (Raguso et al. 1994). The glucose-sparing effect of sebacate seemed to be directly exerted at the 170.106.33.42 intracellular level, because insulin secretion was not stimulated (no modifications of plasma C-peptide values were shown), nor was sebacate plasma clearance influenced by hyperinsulinaemia. , on Dodecanedioic acid (C12) is a saturated aliphatic DA with twelve C atoms which, 28 Sep 2021 at 04:31:38 contrary to shorter chain DA, presents a very low urinary excretion (Mingrone et al. 1994; Bertuzzi et al. 1995). The aim of the present study was to investigate the tissue uptake of C12 and the effect of its acute administration on glucose kinetics. MATERIALS AND METHODS , subject to the Cambridge Core terms of use, available at Experimental protocol Three groups of male Wistar rats (average body weight (BW) 200 g, fasted for about 8 h) were injected intraperitoneally with 0.8 mmol/kg BW of C12 (in the form of the disodium salt) as a bolus (n 35, group A, and n 35, group B), or of an equal volume of physiological salt solution (n 30, group C). Five additional rats, injected with C12 as described earlier, were allocated to individual metabolic cages in order to collect 24 h urine samples. Blood samples were drawn by cardiac puncture, one for each animal of group A, at different times after the bolus in the range from 5 to 90min. The animals were then anaesthetized with diethyl ether and immediately killed by cervical dislocation. No anaesthetic was used before any experimental manoeuvre. All rats in the B and C groups received, immediately after injection of C12 or saline, an intravenous bolus (tail vein injection) of 139 kBq D-[U-'4C]glUCOSe (Amersham, Bucks; specific activity 11248 MBq/ mmol). Blood samples were drawn, in the same fashion as in group A, between 5 and https://www.cambridge.org/core/terms 120 min after the bolus. Heparinized blood samples were immediately centrifuged at 4000g and plasma was frozen at -20" until analysis. Analytical methods To measure the tracer concentration in plasma, 0-5ml plasma was put in a glass test-tube cooled on ice. Ice-cold 700 ml/l acetone in water (4 ml) was added to the tube and stirred gently. The sample was centrifuged at approximately 6000 rev./min for 10 min. Plasma extract was dried in a GyroVap apparatus (Howe model GV1, Gio. De Vita, Rome, Italy) operating at 60", coupled with a vacuum pump and a gas trap FTS System (Stone Ridge, . https://doi.org/10.1079/BJN19970126 New York, USA). The extract was resuspended in acetone-water (4 : 1, v/v) and deposited on silica gel Fluka G plates (200 x 200 mm) with binder for TLC, impregnated with 0.2 M- sodium acetate. The layers were dried for 24 h at room temperature and activated at 1 10" for 1 h. Extracts were eluted using acetone-water (9 : 1, v/v> as the solvent system. With this method, the RF of glucose was equal to 25%. The spots were detected by Downloaded from DODECANEDIOIC ACID KINETICS 145 https://www.cambridge.org/core naphthoresorcinol-H2S04 reagent, and those corresponding to glucose were scraped out of the plates and put in vials, adding 0.5 ml water and 10 ml Insta-FluorTM(Packard, Downers Grove, IL, USA). I4C radioactivity was measured by a p-scintillation counter (Model 1600TR Canberra-Packard, Meriden, CT, USA). Quenching was checked by the internal standard method. In order to measure the total recovery of D-[U-'4C]glUCOSe, amounts ranging from 33.67Bq to 336.7Bq were added to 1 ml samples of pooled plasma. At the smallest labelled glucose concentration the recovery calculated on ten replicate . IP address: experiments ranged between 90 and 95%. This type of TLC method allows separation of many kinds of carbohydrates with good sensitivity (Zweig & Sherma, 1972). Plasma glucose concentration was determined on a Beckman Glucose Analyzer I1 170.106.33.42 (Beckman Instruments, Fullerton, PA, USA). The concentration of total (bound plus free) dodecanedioic acid in plasma and urine was measured by HPLC as previously described (Mingrone et al. 1991). Plasma albumin concentration was determined by a standard colorimetric method. , on 28 Sep 2021 at 04:31:38 Kinetics of dodecanedioic acid The kinetics of C12 were described by a one-compartment model. Since C12 binds to plasma albumin, the total plasma C12 concentration, ct (molA), can be represented by the , subject to the Cambridge Core terms of use, available at sum of a free C12 fraction, c (mol/l), and a bound C12 fraction. Previous results (Mingrone et al. 1994) indicate that binding follows an apparent one-step kinetic, so that the total plasma C12 concentration can be written KC c,=c+nA- 1 + Kc' where n is 1.57 and is the mean number of binding sites per albumin molecule and K is 6.8 x 1031/m01 and is the association constant. In equation (l), A denotes the mean albumin concentration in the plasma of the rats under study, which was equal to 48 gA.
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